REGULATION OF THE MACROPHAGE INFLAMMATORY PHENOTYPE IN ARDS

ARDS 中巨噬细胞炎症表型的调节

• 批准号: 10094230
• 负责人: John W Christman
• 金额: $ 56.37万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10094230
• 关键词: ADP ribosylation; Ablation; Acute Lung Injury; Address; Adoptive Transfer; Adult Respiratory Distress Syndrome; Affect; Affinity; Alveolar; Alveolar Macrophages; Antibiotics; Apoptosis; Attenuated; Binding; Bleomycin; Blood capillaries; Bone Marrow; Calcineurin; Caring; Cells; Chimera organism; Clinical; Critical Illness; Data; Development; Endothelium; Family; Functional disorder; Gene Expression; Generations; Genetic; Health; Human; Hypoxemia; Inflammation; Inflammatory; Intensive Care Units; Knockout Mice; Laboratories; Life; Ligands; Liquid substance; Lung; Measures; Mediating; Mediator of activation protein; Methods; Molecular; Molecular Target; Morbidity - disease rate; Mus; Outcome; Oxygen; Pathogenesis; Patient-Focused Outcomes; Peptides; Permeability; Pharmacological Treatment; Pharmacology; Phenotype; Pneumonia; Prevention approach; Production; Proteolysis; Public Health; Pulmonary Edema; Pulmonary Inflammation; Recovery; Regulation; Research; Role; TNF gene; TNF-related apoptosis-inducing ligand; Therapeutic Agents; Ventilator; base; cecal ligation puncture; clinically relevant; cytokine; design; experimental study; high risk; improved; improved outcome; inflammatory lung disease; inhibitor/antagonist; macrophage; member; mortality; mouse model; neutrophil; novel; novel strategies; nuclear factors of activated T-cells; patient population; pre-clinical; preclinical efficacy; prevent; promoter; pulmonary function; reconstitution; recruit; response; therapeutically effective; transcription factor; transcription factor NF-AT c3; ventilation

PROJECT SUMMARY Acute Lung Injury (ALI) and Acute Respiratory Distress Syndrome (ARDS) are neutrophilic inflammatory lung diseases with unacceptably high rates of morbidity and mortality. Our supportive data indicate that genetic ablation and pharmacologic inhibition of NFATc3 is beneficial in terms of tighter pulmonary barrier function, decreased cytokine release and neutrophilic inflammation, improved arterial oxygenation, and better survival in mouse models of ARDS/ALI. Based on these and other clinically relevant supportive data, we posit that NFATc3 activation in macrophages induces inflammatory gene expression that mediates the initiation, intensity and duration of ALI/ARDS. We propose three specific aims to address this hypothesis: Specific Aim 1: To identify the mechanism(s) by which NFATc3 deletion protects against ALI/ARDS. We will employ genetically defined mice that globally lack NFATc3 subjected to a well-defined murine model of ALI/ARDS. We will also utilize established methods of NFATc3 KO/WT bone marrow chimera mice, macrophage passive adoptive transfer of lung macrophages, and we will construct novel conditional macrophage selective NFATc3-/- mice to identify the impact of macrophage NFATc3 deficiency on the generation of pulmonary edema, arterial oxygen saturation, and mortality. Specific Aim 2: To determine the mechanisms by which macrophage NFATc3 activation contributes to the development of permeability pulmonary edema. In order to determine how macrophages contribute to the pulmonary edema and severe hypoxemia in ALI/ARDS, we propose to examine the role of NFATc3 regulation of macrophage expression of TNF-related apoptosis-inducing ligand (TRAIL), which inhibits alveolar fluid clearance (AFC), in addition to lung macrophage production of mediators that disrupt barrier function of pulmonary microvascular endothelium. Specific Aim 3: To determine the efficacy of a novel cell permeable peptide NFAT inhibitor in protecting against ALI/ARDS-like pathophysiology in mouse models. Our group has developed a novel NFAT inhibitor, CP9-ZIZIT, that has an impressively low Kd value of 2.6 nM, better cell permeability, higher binding affinity, and improved stability against proteolysis than other available agents. We expect to show that treatment with CP9-ZIZIT alone or in combination with a PARP1 inhibitor will attenuate pulmonary inflammation and alveolar capillary barrier disruption. We expect that this research will provide valuable preclinical data that blocking NFATc3 activation in macrophages is an effective therapeutic agent for preventing ARDS in a high-risk patient population and improving the clinical outcome of patients suffering from moderate and severe ARDS/ALI.

项目摘要 急性肺损伤（ALI）和急性呼吸窘迫综合征（ARDS）是嗜中性炎性肺 发病率和死亡率率高的疾病。我们的支持性数据表明遗传 NFATC3的消融和药理抑制在更紧密的肺屏障功能方面是有益的 细胞因子释放和中性粒细胞炎症减少，动脉氧合的改善以及更好的生存率 ARDS/ALI的鼠标模型。基于这些和其他临床相关的支持数据，我们认为 巨噬细胞中的NFATC3激活诱导介导开始的炎症基因表达， ALI/ARDS的强度和持续时间。我们提出了三个特定的目的来解决这一假设： 特定目的1：确定NFATC3删除保护ALI/ARDS的机制。 我们将采用遗传定义的小鼠，该小鼠缺乏经受明确定义的鼠模型的NFATC3 Ali/ards。我们还将利用NFATC3 KO/WT骨髓小鼠的既定方法， 巨噬细胞被动的肺巨噬细胞的收养转移，我们将构建新颖的条件 巨噬细胞选择性NFATC3 - / - 小鼠，以识别巨噬细胞NFATC3缺乏症的影响 肺水肿的产生，动脉氧饱和度和死亡率。 特定目的2：确定巨噬细胞NFATC3激活有助于的机制 渗透性肺水肿的发展。为了确定巨噬细胞如何贡献 ALI/ARDS中的肺水肿和严重低氧血症，我们建议检查NFATC3的作用 调节TNF相关凋亡诱导配体的巨噬细胞表达（TRAIL），该配体抑制 除了破坏障碍的肺巨噬细胞的产生外，肺泡流体清除率（AFC） 肺微血管内皮的功能。 特定目的3：确定新型细胞可渗透肽NFAT抑制剂保护的功效 针对小鼠模型中的ALI/ARDS样病理生理学。我们的小组开发了一个新颖的NFAT 抑制剂CP9-Zizit，其kD值为2.6 nm，较高的细胞渗透性，更高的结合 与其他可用药物相比，亲和力和针对蛋白水解的稳定性提高。我们希望表明 单独或与PARP1抑制剂结合使用CP9-ZizIT治疗将减弱肺 炎症和肺泡毛细管屏障破坏。 我们希望这项研究将提供有价值的临床前数据，以阻止NFATC3激活 巨噬细胞是预防高风险患者人群ARDS的有效治疗剂 并改善患有中度和重度ARD/ALI患者的临床结果。